Dehydrogenase variants and polynucleotides encoding same

Abstract

The present invention relates to 3-hydroxypropionate dehydrogenase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

What is claimed is: 
     
       1. A 3-HPDH variant, comprising a substitution at one or more positions corresponding to positions 9, 31, 32, 33, 34, 35 and 36 of SEQ ID NO: 2;
 wherein the variant has at least 80% sequence identity to SEQ ID NO: 2, 4, or 6; and 
 wherein the variant has 3-HPDH activity. 
 
     
     
       2. The variant of  claim 1 , comprising a substitution at a position corresponding to position 9 of SEQ ID NO: 2. 
     
     
       3. The variant of  claim 1 , comprising a Gly at a position corresponding to position 9 of SEQ ID NO: 2. 
     
     
       4. The variant of  claim 1 , comprising a substitution at a position corresponding to position 31 of SEQ ID NO: 2. 
     
     
       5. The variant of  claim 1 , comprising an Asp or Glu at a position corresponding to position 31 of SEQ ID NO: 2. 
     
     
       6. The variant of  claim 1 , comprising a substitution at a position corresponding to position 32 of SEQ ID NO: 2. 
     
     
       7. The variant of  claim 1 , comprising a Leu at a position corresponding to position 32 of SEQ ID NO: 2. 
     
     
       8. The variant of  claim 1 , wherein the variant comprises or consists of SEQ ID NO: 7, 8, 9, 10, 11, 12, 13, 14, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 80, or 81. 
     
     
       9. The variant of  claim 1 , wherein the variant has increased specificity for NAD(H) compared to NADP(H). 
     
     
       10. A polypeptide having 3-HPDH activity, wherein the polypeptide has sequence identity to SEQ ID NO: 2, 4, or 6; and wherein the polypeptide has increased specificity for NAD(H) compared to NADP(H). 
     
     
       11. The polypeptide of  claim 10 , wherein the polypeptide has at least 90% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       12. The polypeptide of  claim 10 , wherein the polypeptide has at least 95% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       13. The polypeptide of  claim 10 , wherein the polypeptide has at least 98% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       14. The polypeptide of  claim 10 , wherein the polypeptide has greater than 10-fold specificity for NAD(H) compared to NADP(H). 
     
     
       15. The 3-HPDH variant of  claim 1 , wherein the variant has at least 90% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       16. The 3-HPDH variant of  claim 1 , wherein the variant has at least 95% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       17. The 3-HPDH variant of  claim 1 , wherein the variant has at least 98% sequence identity to SEQ ID NO: 2, 4, or 6. 
     
     
       18. The 3-HPDH variant of  claim 9 , wherein the variant has greater than 10-fold specificity for NAD(H) compared to NADP(H). 
     
     
       19. The 3-HPDH variant of  claim 1 , wherein the variant comprises substitutions at positions corresponding to positions 9, 31, and 32 of SEQ ID NO: 2. 
     
     
       20. The 3-HPDH variant of  claim 1 , wherein the variant comprising a Gly at a position corresponding to position 9 of SEQ ID NO: 2; an Asp or Glu at a position corresponding to position 31 of SEQ ID NO: 2; and a Leu at a position corresponding to position 32 of SEQ ID NO: 2.